Quantitative and qualitative studies on bacterial flora of hybrid tilapia (Oreochromis niloticus × O. aureus) cultured in earthen ponds in Saudi Arabia

The bacterial flora of the rearing pond water and sediment as well as the gills and intestine of healthy hybrid tilapia cultured in Saudi Arabia was estimated quantitatively and qualitatively, the isolates being identified at genus or species level. Total viable counts of bacteria (measured as colony‐forming units, cfu) were in the range 5.6 ± 0.8 × 10³ to 2.4 ± 1.2 × 10⁴ cfu mL^?1 in pond water; 9.3 ± 1.1 × 10⁶ to 1.9 ± 1.5 × 10⁸ cfu g^?1 in sediment; 7.1 ± 0.7 × 10⁵ to 8.7 ± 1.1 × 10⁶ cfu g^?1 in the gills of tilapia; and 3.4 ± 1.8 × 10⁶ to 5.8 ± 0.4 × 10⁷ cfu g^?1 in the intestine of tilapia. In total, 15 bacterial genera and 18 species were identified. Pond water and sediment bacteria reflected the bacterial composition in the gills and intestine of tilapia. In contrast to gill bacteria, more diversification was observed in intestinal bacteria. Corynebacterium urealyticum, Shewanella putrefaciens and Aeromonas hydrophila predominated in all samples. In pond water, C. urealyticurn, S. putrefaciens, A. hydrophila, Flavobacterium sp. and Pseudomonas sp. were the most predominant bacterial species (prevalence > 10%), whereas A. hydrophila, C. urealyticum, S. putrefaciens and Escherichia coli were predominant in pond sediment, and C. urealyticum, S. putrefaciens and A. hydrophila were predominant in both the gills and intestine of tilapia.     

The histopathology of carp, Cyprinus carpio L., exposed to microcystins by gavage, immersion and intraperitoneal administration

Carp liver, gills, intestine, kidneys, heart and spleen were studied by histology after the fish were exposed to microcystins by gavage, immersion and intraperitoneal administration. Intraperitoneal inoculation with microcystins caused necrosis or dose- dependent degeneration in the liver, gills and kidneys. Gavaging with microcystins caused changes in the histopathology of the liver and gills. Cellular degeneration and necrosis occurred in the liver, gills and kidneys when carp were introduced to a tank containing 1.7 μg ml^?1 of microcystins. Lesions were not observed in the heart, spleen or intestines from any of the treated carp. Microcystins administered by intraperitoneal injection at a concentration of 50 μg kg^?1 were lethal to all fish within 8 h, while gavaging with 250 μg kg^?1 of microcystins caused minimal damage in the tissues studied.     

Aerobic Bacterial Flora of Common Carp (Cyprinus carpio L.) Cultured in Earthen Ponds in Saudi Arabia

ABSTRACT

Quantitative and qualitative estimation of bacterial flora present in pond water, sediments, gills, and intestine of healthy common carp Cyprinus carpio cultured in Saudi Arabia were performed and identified to species level where possible. Mean total viable bacterial counts in pond water ranged from 1.2?±?2.9?×?10⁴ to 2.5?±?3.5?×?10⁵ cfu/mL; in sediments, 9.3?±?2.1?×?10⁷ to 2.7?±?3.5?×?10⁹ cfu/g; in gills filaments, 4.3?±?2.9?×?10⁶ to 1.6?±?3.9?×?10⁷ cfu/g; and in intestine, 8.7?±?4.1?×?10⁹ to 5.4?±?3.2?×?10¹⁰ cfu/g. Gram-negative rod-shaped bacteria dominated (76%) the populations. In total, 12 bacterial genera and 15 species were identified. Pond water and sediment bacteria had the reflection on bacterial composition of gills and intestine of carp. Intestinal bacteria showed more diversification in contrast to gill bacteria. Aeromonas hydrophila, Shewanella putrefaciens, Vibrio cholerae, Corynebacterium urealyticum, Vibrio vulnificus, Vibrio sp., and Staphylococcus sp. were the common bacteria in all the populations. In pond water and carp intestine, A. hydrophila, S. putrefaciens, V. cholerae, and C. urealyticum were the most dominant bacteria (prevalence ≥ 10%) where pond sediments and the carp gills experienced with more one dominant bacterium V. vulnificus. Only the A. hydrophila covered one fourth (25%) of the total bacterial populations.     

Effect of dietary protein-energy levels and fish initial sizes on growth rate, development and production of Nile tilapia, Oreochromis niloticus L.

The effects of dietary protein‐energy levels on the growth rate, proximate composition and production were examined in Nile tilapia, Oreochromis niloticus, at two starting weights (22.9 and 39.8 g) reared in concrete ponds for 180 days. The highest weight gain (183.1 g) was obtained with fish fed a 30% protein and 10.5 kJ g^?1 diet for the small initial size and 180.2 g for a diet containing 25% protein and 12.6 kJ g^?1 for the large initial size. Dressed yields (edible mass) and fillets increased to 56.9% and 52.5% in fish fed diet with 25% protein and 10.5 kJ g^?1 at the initial size of 22.9 g, while fish started at 39.8 g exhibited the best values (56.5% and 52.1%) when fed the 30% protein and 10.5 kJ g^?1 diet. Proximate composition of soft tissue (wet weight basis) in small fish was significantly influenced by dietary protein‐energy levels. Protein was 26.1±0.3% in fish fed the high protein (30%) and low energy (10.5 kJ g^?1 diet), while lipid content was 6.4±0.3% at diet containing 20% protein and 14.7 kJ g^?1 diet. Large initial size fish fed the diet with 25% protein and 14.7 kJ g^?1 had the highest body protein (32.0±0.4%) and lowest lipid content (2.2±0.3%). Feed conversion ratio (FCR) and protein efficiency ratio varied with different dietary protein‐energy levels and initial fish sizes. Feed conversion ratio increased with increasing protein and decreasing energy level in the diet, and values in small fish were higher than values in large fish. Protein efficiency ratio decreased with increasing dietary protein level and decreasing energy level. The maximum total production (7.6 tons feddan^?1) was with dietary high protein (30%) and low energy (10.5 kJ g^?1) for small‐sized fish, while large initial fish had the highest production (3.7 tons feddan^?1) when fed the 25% protein and 12.6 kJ g^?1 diet energy. Starting with 22.9 g fish was more advantageous than the initial size of 39.8 g for rearing Nile tilapia. Small fish required a high‐protein and low‐energy diet, whereas large fish required a low‐protein and high‐energy diet to achieve highest production.     

Effect of esterification on the absorption of astaxanthin in rainbow trout, Oncorhynchus mykiss (Walbaum)

Gastrointestinal and serum absorption of astaxanthin was studied in rainbow trout, Oncorhynchus mykiss (Walbaum) (217 ± 2 g) fed diets supplemented with either esterified astaxanthin (from Haematococcus pluvialis) or free astaxanthin (synthetic, as 8% w/w beadlets) at similar levels (50 mg kg^?1). After 56 days of feeding, there was a significant difference (P = 0.0582) between steady‐state serum astaxanthin concentrations for fish fed free (2.0 ± 0.3 μg mL^?1) or esterified astaxanthin (1.3 ± 0.1 μg mL^?1) at the 90% confidence level. However, following ingestion of a single meal supplemented with free or esterified astaxanthin, the rates of astaxanthin absorption into serum were not significantly different (P > 0.1) (0.8 ± 0.2 µg mL^?1 h^?1 and 1.0 ± 0.4 µg mL^?1 h^?1 respectively). In fish fed both free or esterified astaxanthin, higher absorption (P < 0.05) of astaxanthin by the ileal (0.8 ± 0.14 μg g^?1 and 0.9 ± 0.15 μg g^?1 respectively) compared with the posterior (0.2 ± 0.01 μg g^?1 and 0.3 ± 0.14 μg g^?1 respectively) intestine was recorded. This confirmed the role of the anterior intestine in carotenoid absorption. Non‐detectable levels of esters in digesta taken from the hind intestine suggest the anterior intestine is also the primary region for ester hydrolysis.     

Faecal coliforms in pond water, sediments and hybrid tilapia Oreochromis niloticus×Oreochromis aureus in Saudi Arabia

Total bacterial load, total coliforms faecal coliforms in pond water, sediment, intestine of hybrid tilapia Oreochromis niloticus×Oreochromis aureus and pigeon Columba livia faeces were investigated monthly over a period of 1 year from July 1999 to June 2000. Fish were collected randomly by a cast net. Samples were analysed for coliforms using the multiple‐tube fermentation technique. Results showed total viable bacterial counts in the pond water, sediment, intestine of tilapia and pigeon faeces ranging from 1.8±0.9×10² to 6.0±1.2×10⁴ cfu mL^?1, 3.2±1.2×10⁵ to 2.8±1.5×10⁷ cfu g^?1, 8.2±1.6×10⁵ to 9.9±1.5×10⁷ cfu g^?11.0±0.4×10⁷to9.7±0.2×10⁹ cfu g^?1respectively. The most probable number (MPN) of coliforms and faecal coliforms ranged from 287±12 to ≥1600±0 100 mL^?1 in pond water; the MPN ranges for sediment, tilapia intestine and pigeon faeces were 257±29 to ≥1100±0 g^?1, 237±46 to ≥1100±0 g^?1 and 403±98 to ≥1100±0 g^?1 respectively. The abundance of normal bacteria coliforms was greater in the warm months than in the cold months. Ground water was free from any sort of coliform organisms, and there were no sources of human faecal matter in the pond. So, it is clear that faecal coliforms from pigeon faeces significantly contaminated (P<0.05) the ponds and tilapia intestines. Escherichia coli was the only coliform organism found in pond water, sediment, intestine of tilapia and pigeon faeces.     

The fate of chlortetracycline residues in a simulated chicken–fish integrated farming systems

Two experiments were conducted at the Asian Institute of Technology, Pathumthani, Thailand to investigate the fate of chlortetracycline (CTC) residue in chicken manure and its effect on integrated chicken–fish farming system. During the first experiment, broiler chickens were raised and CTC residues in their manure were analysed. Chicken fed diets containing 0, 50, 200 and 800 CTC mg kg^?1 had CTC residue levels of 0, 0.9, 3.8 and 6.5 CTC ng g^?1. Once the diet containing CTC was withdrawn, CTC in the manure dropped to negligible amounts (0, 0, 0.2 and 0.5 CTC ng g^?1) within 1 day. Integrated chicken–fish farming systems were simulated during the second experiment to determine the fate of antibiotic residues in chicken manure in aquaculture environment. Chickens were fed a CTC‐free diet and a feed containing CTC at 200 mg kg^?1. Ten 4 m³ square concrete tanks (2 × 2 × 1 m) were used for the experiment. Five tanks were fertilized with CTC‐contaminated manure and the remaining five tanks were fertilized with CTC‐free manure at a rate of 100 kg dry matter ha^?1 day^?1. Sex‐reversed Nile tilapia (Oreochromis niloticus) was stocked at 12 fish tank^?1 on the 14th day after chicken manure application. The immuno‐radio microbial receptor assay (Charm II test) revealed that edible fish muscle, fish intestinal tract and sediment were contaminated by CTC at rates of 7.21, 22.104 and 1.788 ng g^?1, respectively, after 45 days. Chlortetracycline was detected on day 20 in the water column and gradually increased from 0.26 to 12.13 ng g^?1. Chlortetracycline residues were not detected in fish or the aquatic environment of the CTC‐free treatment. The results demonstrate the potential for antibiotic residue accumulation in fish and aquatic environment when CTC‐contaminated chicken manure is used for pond fertilization.     

Chronic effect after acute exposure to commercial petroleum fuels on physiological status of Nile tilapia, Oreochromis niloticus (L.)

Quantitative and qualitative analyses of bacterial flora associated with pond water, gills, and intestine of polycultured healthy common carp (Cyprinus carpio) and African catfish (Clarias gariepinus) were carried out and identified to species level where possible. Total viable bacterial counts in the pond water ranged from 9.2?±?5.5?×?10³ to 6.6?±?5.1?×?10⁴ colony-forming units (cfu)/mL; in the gill filaments of carp and catfish, 3.3?±?3.8?×?10⁶ to 7.9?±?5.6?×?10⁶ and 1.1?±?4.6?×?10⁵ to 2.3?±?5.2?×?10⁶ cfu/g, respectively; and in the intestine of carp and catfish, 1.4?±?2.9?×?10¹⁰ to 1.7?±?6.0?×?10¹¹ and 2.7?±?3.4?×?10¹⁰ to 1.0?±?4.5?×?10¹¹ cfu/g, respectively. Gram-negative rod-shaped bacteria dominated the populations: 90% in carp, 89% in catfish, 80% in water, and 86% in the total populations. Altogether, 14 bacterial species of 10 genera were identified in total populations. Pond water bacteria had a reflection on the bacterial composition of the gills and intestine of carp and catfish. Aeromonas hydrophila, Shewanella putrefaciens, Vibrio cholerae, Staphylococcus sp., and Vibrio vulnificus appeared as the common bacteria in the populations, where the first three were highly significantly abundant (P?<?0.0001). Moreover, A. hydrophila was the most significantly dominant bacteria (32%; P?<?0.005) among the total populations. Pantoea sp. and Pasteurella pneumotropica were present only in carp and catfish, respectively, but Corynebacterium urealyticum and Micrococcus sp. were present only in pond water.     

Effect of sublethal toxicity of some pesticides on growth parameters, haematological properties and total production of Nile tilapia (Oreochromis niloticus L.) and water quality of ponds

The effects of sublethal doses of dimethoate (20, 10 and 5 mg L^?1) and malathion (2.0, 1.0 and 0.5 mg L^?1) on growth parameters, haematological properties and total production of Nile tilapia (12.0 cm length and 40.0 g weight) and water quality were studied in fibreglass tanks (3 m²) for 24 weeks. Results showed that malathion compounds were more toxic than dimethoate. The sublethal toxicity of pesticides decreased plankton abundance and water quality in fish ponds. In comparison with controls, sublethal levels of these pesticides led to a significant decrease (P<0.05) in final body weight, specific growth rate and normalized biomass index of fish. The survival rate of Nile tilapia decreased with increasing concentrations of pesticides. Feed utilization (total food consumed, feed conversion ratio and protein efficiency ratio) varied with pesticide treatments. The erythrocyte count, haematocrit value and haemoglobin content of Nile tilapia were slightly decreased with increasing concentrations of pesticides. In addition, serum protein decreased, while serum glucose and lipid increased. Further, glycogen, protein and lipid in fish muscle gradually decreased with increased pesticide concentrations. Pesticide residues in the liver, gills and muscles of fish increased with increased pesticide concentrations in fish ponds. Their bioaccumulation in the liver was higher than in gill or muscle, which had the lowest residues for these pesticides. On the other hand, total production, net returns and profitability of reared fish decreased with increasing of concentrations of pesticides. Generally, sublethal concentrations of two pesticides in aquatic areas led to degradation of water quality and fish production in ponds.     

Acute and chronic effects of aqueous ammonia on marbled spinefoot rabbitfish,Siganus rivulatus (Forsskål 1775)

Ammonia is a metabolite of aquatic organisms which might reach deleterious levels in intensive fish farms. The aim of the present study was to determine median lethal concentrations (96‐h LC₅₀) of total ammonia nitrogen (TA‐N) on marbled spinefoot rabbitfish (Siganus rivulatus) and chronic effects of TA‐N on survival, growth and behaviour of juvenile rabbitfish over a 50 day period. In the first experiment, fish were exposed to 0, 2, 4, 6, 8, 10, 12, 14, 16, 18 and 20 mg L^?1 TA‐N for 96 h and survival evaluated. In the second experiment, 12 fish were stocked per 50‐L tank and treated with one of 0, 2, 4, 6, 8, 10 and 12 mg L^?1 TA‐N with three replicate tanks per treatment. Survival and growth were determined and histopathological alterations of gills due to chronic ammonia exposure were studied by light and electron microscopy. The 96‐h LC₅₀ values were 16–18 mg L^?1 TA‐N. In the chronic exposure experiment, fish reared in water with 0 mg L^?1 TA‐N had 100% survival and had 50% weight increase in 50 days. Fish at 2 and 4 mg L^?1 TA‐N all died whilst fish in 6, 8, 10 and 12 mg L^?1 TA‐N survived and grew albeit less than in treatment 0 mg L^?1. Gills from ammonia treated fish displayed severe histological and ultrastructural alterations including hyperplasia, hypertrophy and fusion of secondary lamellae, aneurysms and presence of pleomorphic altered cells. Chronic exposure to ammonia is deleterious to marbled spinefoot rabbitfish and low concentrations of ammonia appear to kill the fish in <50 days whilst fish can survive for more than 50 days at concentrations between 6 and 12 mg L^?1 TA‐N.     

Oral administration of bovine lactoferrin inhibits bacterial infection in tilapia and elevates survival after bacterial infection: an examination of its immune-modulating properties

It was previously reported that the oral administration of lactoferrin (LF) provides antimicrobial activity in animals against bacterial and viral infections and is involved in immunomodulatory properties. In this report, a hybrid strain of Oreochromis nilotica (male) × O. mossambicus (female) was fed homemade diets with or without supplementation with 10, 50, 100, and 150 mg bovine LF g^?1 feed for 60 days. After dietary treatment, the experimental fish were infected with Streptococcus agalactiae. LF supplementation resulted in a significantly higher survival rate and suppression of bacterial growth at 24–96 h in the liver, spleen, kidneys, eyes, and gills. A reduction in peroxidase activity was followed by a similar reduction in the peroxidase content of leukocytes at 24 h as analyzed by spectrophotometry. Respiratory burst (RB) activity was detected regardless of the time at which LF was administered to fish in relation to the bacterial infection. A beneficial effect of LF on RBs was also detected before bacterial challenge (with 100 mg LF/g) and 24 h after bacterial challenge with 150 mg/g. The results obtained validate LF’s beneficial effects on RBs by phagocytes in tilapia, which was only shown at 48 h after supplementation with 100 mg bovine LF g^?1 feed after the bacterial infection. Moreover, after oral administration of LF, it was detected in the mucosa of the small intestines of tilapia. In conclusion, we demonstrate a reduction in fish mortality after the oral administration of LF, and we examined its immunomodulatory properties in tilapia.     

Effects of dietary phenylalanine on growth,digestive and brush border enzyme activities and antioxidant capacity in the hepatopancreas and intestine of young grass carp (Ctenopharyngodon idella)

This study investigated the effects of phenylalanine on growth, digestive and absorptive ability and antioxidant status of young grass carp (Ctenopharyngodon idella). Young grass carp were fed diets containing 3.4 (basal diet), 6.1, 9.1, 11.5, 14.0 and 16.8 g phenylalanine kg^?1 diet with a fixed of 10.7 g tyrosine kg^?1 diet for 8 weeks. Percent weight gain (PWG), feed efficiency and feed intake of fish were the lowest in fish fed the basal diet (P < 0.05). Trypsin, lipase and amylase activities in the hepatopancreas, and antioxidants including glutathione contents and glutathione reducase activities in the hepatopancreas and intestine were all the highest in fish fed 11.5 g phenylalanine kg^?1 diet (P < 0.05). Trypsin, chymotrypsin and amylase activities in whole intestine, and creatine kinase, Na⁺, K⁺‐ATPase and alkaline phosphatase activities in the proximal intestine, and superoxide dismutase activities in the hepatopancreas and intestine were all the highest when phenylalanine at level of 9.1 g kg^?1 diet (P < 0.05). In conclusion, phenylalanine improved growth, digestive and absorptive ability, and antioxidant capacity of young grass carp. The phenylalanine requirement of young grass carp (256–629 g) based on PWG was 10.4 g kg^?1 diet or 3.44 g 100 g^?1 protein.     

Efficacy and toxicity of oxidative disinfectants for the removal of gill amoebae from the gills of amoebic gill disease affected Atlantic salmon (Salmo salar L.) in freshwater

Amoebic gill disease (AGD) of Atlantic salmon is treated commercially by bathing affected fish in freshwater. Recently, the efficacy of freshwater bathing has been questioned, and the aim of this study was to examine the potential for improving bathing efficacy using additives to the freshwater bath. AGD‐affected Atlantic salmon were bathed in 350 L tanks containing oxygenated freshwater to which chlorine dioxide (0–50 mg L^?1), chloramine‐T (0–50 mg L^?1) or hydrogen peroxide (0–100 μL L^?1) was added. Before and following a 3‐h exposure to the freshwater and chemical additive, the gills were removed from a sub‐sample of fish and the number of live amoebae on the gills were counted and smears made for confirmation of the presence of Neoparamoeba pemaquidensis, the causative agent of AGD. Following a further 3‐h exposure, a sub‐sample of fish was bled from the caudal vein and the gills were removed for histological examination. Chlorine dioxide and chloramine‐T at 25–50 and 10–50 mg L^?1, respectively, reduced the number of amoebae on the gills by approximately 50% compared with pre‐exposure numbers. The results from hydrogen peroxide treatment were equivocal and the toxicity of hydrogen peroxide was high. The toxicity of chlorine dioxide varied with freshwater hardness and/or suspended solid load, whereas chloramine‐T toxicity was low, with mortalities attributable only to elevated temperatures at the highest concentration tested. In conclusion, chlorine dioxide and chloramine‐T show promise as potential freshwater additives for the improved removal of N. pemaquidensis and possibly, other amoebae from the gills of commercially farmed Atlantic salmon.     

Dietary uptake of dioxins (PCDD/PCDFs) and dioxin-like PCBs in Atlantic salmon (Salmo salar)

Atlantic salmon (Salmo salar) were fed graded levels of dioxins (polychlorinated dibenzo‐p‐dioxins and polychlorinated dibenzofurans) and dioxin‐like polychlorinated biphenyls (DLPCBs) in their diets for 7 months. The dioxin and DLPCB concentrations in both fillet and whole body of salmon increased with increasing dietary exposure. DLPCBs transferred more efficiently from the feed to edible flesh of salmon than dioxins, and contributed a higher proportion to the total toxic equivalents (TEQ). At the end of the trial, the maximum concentrations of dioxins in fillet and whole fish were 1.9 and 2.3 pg WHO‐TEQ g^?1 fresh weight, respectively. Hence with this feeding period even with the most contaminated feed (4.9 pg WHO‐TEQ g^?1 dw) the dioxin concentrations in salmon did not exceed the maximum level set by the European Commission [4 pg WHO‐TEQ g^?1 (EC 2375/2001)]. The inclusion of DLPCBs in this study provides valuable information for forthcoming risk assessments and the future establishment of maximum limits for these compounds in feed and fish.     

The effects of diet supplemented with Lactobacillus rhamnosus on tissue parameters of rainbow trout,Oncorhynchus mykiss (Walbaum)

This study was carried out to establish the effects of a 6 week treatment with the diet supplemented with L. rhamnosus in concentrations of 10⁷ CFU g^?1 (G1 group) and 10⁸ CFU g^?1 (G2 group) on the condition expressed by condition factors (Fulton's, Clark's and B), intestinal microbiology, haematological, histological and selected antioxidative parameters of rainbow trout. A significantly higher condition factors were found in G1 group indicating that higher concentration of probiotic (10⁸ CFU g^?1) did not result in the better condition. Cholesterol and urea levels were significantly higher in both G1 and G2 groups, albumin in G1 and creatinine in G2 group with respect to control. A significantly higher liver TBARS level was observed in G2 group. The feeding with supplemented probiont apparently changed the resident microbiota. Three weeks after withdrawal of the supplemented feed, the microflora mostly reverted to the control composition, although L. rhamnosus in faecal matter of fish remained inherent. The epithelial structure of the proximal and distal intestine revealed the increased absorptive area in both treated groups, as well as the increase in the mucin‐secreting goblet cells. The L. rhamnosus‐treated groups demonstrated the capacity for the augmentation of the innate host defence.     

Evaluation of optimum dietary protein-to-energy ratio in juvenile olive flounder Paralichthys olivaceus (Temminck et Schlegel)

An 8‐week feeding trial was conducted to estimate the optimum dietary protein to energy (P/E) ratio in juvenile olive flounder Paralichthys olivaceus. Eight experimental diets were formulated with two energy levels and four protein levels at each energy level. Two energy levels of 12.5 and 16.7 kJ g^?1 diets were included at crude protein (CP) levels of 25%, 30%, 35% and 45% with 12.5 kJ g^?1, and CP levels of 35%, 45%, 50% and 60% with 16.7 kJ g^?1. After 1 week of the conditioning period, fish initially averaging 8.1±0.08 g (mean±SD) were randomly distributed into the aquarium as groups of 15 fish. Each diet was fed on a dry‐matter basis to fish in three randomly selected aquariums at a rate of 3–5% of total wet body weight per day for 8 weeks. After 8 weeks of the feeding trial, weight gain (WG), feed efficiency ratio and specific growth rate of fish fed 45% CP with 16.7 kJ g^?1 energy diet were significantly higher than those from the other dietary treatments (P<0.05). WG of fish fed 12.5 kJ g^?1 energy diets increased with the increase of dietary protein levels. However, WG of fish fed 16.7 kJ g^?1 energy diets increased with the increase of dietary protein levels up to 45% CP and then decreased when fish fed 50% and 60% CP diets. Both dietary protein and energy affected protein retention efficiency and energy retention efficiency. Haemoglobin (Hb) of fish fed 35% and 45% CP diets with 12.5 kJ g^?1 energy were significantly high and not different from Hb of fish fed 45% and 50% CP diets with 16.7 kJ g^?1 energy. Haematocrit of fish fed 45% CP diet with 16.7 kJ g^?1 energy was significantly higher than those from fish fed 25% and 30% CP diets with 12.5 kJ g^?1 energy (P< 0.05). Based on the results of this experiment, we concluded that the optimum dietary P/E ratio was 27.5 mg protein kJ^?1 with diet containing 45% CP and 16.7 kJ g^?1 energy in juvenile olive flounder.     

Diversity of siderophore‐producing bacteria isolated from the intestinal tracts of fish along the Japanese coast

This study examined the diversity of siderophore‐producing bacteria in the intestinal tracts of coastal fish in Japan and screened candidate bacterial strains for probiotic use in aquaculture. Of the 2637 bacteria isolated from the 27 fish specimens (13 species) and six environmental samples collected in this study, 266 isolates exhibited the ability to produce siderophores. Siderophore producers were detected in the intestines of 18 of the fish specimens caught (68%) at densities of 2.3 × 10⁴–2.3 × 10⁸ CFU g^?1, in all three seawater samples at 2.0 × 10²–1.3 × 10³ CFU mL^?1 and in all three sand samples at 2.6 × 10¹–2.8 × 10⁴ CFU g^?1. These findings suggest that siderophore‐producing bacteria are widely distributed in the intestinal tracts of coastal fish and their environments. Analysis of 16S rRNA gene sequences revealed that the siderophore producers belonged to 38 species, of which Vibrio splendidus, Vibrio ichthyoenteri and Vibrio crassostreae accounted for 32.7%, 19.5% and 11.3% of the 266 isolates, respectively, suggesting that these bacteria are indigenous to the intestinal tract of coastal fish. Six bacterial species, Enterovibri norvegicus, Photobacterium leiognathi, Photobacterium phosphoreum, Photobacterium rosenbergii, V. crassostrea and Vibrio scophthalmi were identified as possible candidates for use as probionts in fish aquaculture.     

Vibrio alginolyticus infection in Asian seabass (Lates calcarifer,Bloch) reared in open sea floating cages in India

Asian seabass (Lates calcarifer) has been designated as a candidate species for open sea cage culture in India. A case of vibriosis in Asian seabass reared in open sea floating cages is reported. Haemorrhage and ulcer were observed grossly in the diseased fish. The pathogen, identified as Vibrio alginolyticus based on biochemical and molecular characterization, was isolated from liver, gill, kidney, brain and blood. Histological examination of the diseased fish showed congestion, haemorrhage and necrosis in vital organs. The LD₅₀ studies showed that the organism was virulent to Asian seabass, and the LD₅₀ value was 10^3.2 CFU g^?1 fish.     

Nursery rearing of the Asian catfish, Clarias macrocephalus (Günther), at different stocking densities in cages suspended in tanks and ponds

Growth and survival of hatchery‐bred Asian catfish, Clarias macrocephalus (Günther), fry reared at different stocking densities in net cages suspended in tanks and ponds were measured. The stocking densities used were 285, 571 and 1143 fry m^?3 in tanks and 114, 228 and 457 fry m^?3 in ponds. Fish were fed a formulated diet throughout the 28‐day rearing period. Generally, fish reared in cages in ponds grew faster, with a specific growth rate (SGR) range of 10.3–14.6% day^?1, than those in cages suspended in tanks (SGR range 9–11.3% day^?1). This could be attributed to the presence of natural zooplankton (copepods and cladocerans) in the pond throughout the culture period, which served as additional food sources for catfish juveniles. In both scenarios, the fish reared at lower densities had significantly higher SGR than fish reared at higher densities. In the pond, the SGR of fish held at 228 and 457 m^?3 were similar to each other but were significantly lower than those of fish held at 114 m^?3. The zooplankton in ponds consisted mostly of copepods and cladocerans, in contrast to tanks, in which rotifers were more predominant. Per cent survival ranged from 85% to 89% in tanks and from 78% to 87% in ponds and did not differ significantly among stocking densities and between rearing systems. In conclusion, catfish nursery in cages suspended in tanks and ponds is density dependent. Catfish fry reared at 285 m^?3 in tanks and at 114 m^?3 in ponds had significantly faster growth rates than fish reared at higher densities. However, the desired fingerling size of 3–4 cm total length for stocking in grow‐out culture can still be attained at stocking densities of 457 m^?3 in nursery pond and 571 m^?3 in tanks.     

Effects of addition of a bird repellent to fish diets on their growth and bioaccumulation

The effects of adding the nonlethal bird repellent methyl anthranilate (MA), at levels of 100 and 1000 mg kg^?1, to fish feed on the bioaccumulation and growth of juvenile (10 g) hybrid striped bass (Morone chrysops×M. saxatilis) and juvenile (1 g) African cichlid fish Aulonocara jacobfreibergi were investigated under laboratory conditions. The bird repellent did not have any effect on the fish growth or survival over a period of 6 weeks. MA residues at low levels of 11.2±2.6 μg g^?1 were found in lipophilic tissues (liver) of MA‐fed fish. Control fish, which had no MA added to their diet, had a much lower level of 0.6±0.3 μg g^?1 MA in their liver. Fish muscle was found to contain negligible MA residues, while the outer body surface mucus did not contain any MA. Following a 6‐week depuration period, during which the previously MA‐fed hybrid striped bass were fed a feed to which no MA was added, the levels of MA residues detected were reduced by one order of magnitude.